Application method of TAT-CRYAB fusion protein in intestinal inflammation

An intestinal inflammation, fusion protein technology, applied in DNA/RNA fragments, peptide/protein components, chemical instruments and methods, etc., can solve problems such as rare clinical treatment of diseases, and achieve the improvement of disease activity, protection of integrity, inhibition of Effects of Inflammatory Response

Pending Publication Date: 2019-12-13
杜鹏 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, clinical treatment of diseases using this technology is relatively rare

Method used

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  • Application method of TAT-CRYAB fusion protein in intestinal inflammation
  • Application method of TAT-CRYAB fusion protein in intestinal inflammation
  • Application method of TAT-CRYAB fusion protein in intestinal inflammation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] A method for applying TAT-CRYAB fusion protein in intestinal inflammation, specifically comprising the following steps:

[0050] S1. Construction of pET28a-TAT-CRYAB recombinant plasmid; the construction method is as follows:

[0051] S1.1. Purchase the pET28a plasmid and prepare the pET28a vector digestion system: pET28a vector, NheI, 10xFastdigestBuffer and DEPC water. The details of the pET28a vector digestion system are as follows:

[0052]

[0053] Then prepare the enzyme digestion system, bathe in a 37°C water bath for 1 hour, and then glue it back with the above steps;

[0054] S1.2, pET28a-TAT plasmid connection;

[0055] First, anneal the front and back primers of TAT, and connect after annealing. The system is: 10×T4buffer, T4 ligase, pET28a digestion product and TAT sequence annealing product.

[0056]

[0057] After the system is prepared, let it stand at room temperature for 1 hour;

[0058] S1.3, then transform the pET28a-TAT plasmid, plate, shake...

Embodiment 2

[0079] A method for applying TAT-CRYAB fusion protein in intestinal inflammation, specifically comprising the following steps:

[0080] S1. Construction of pET28a-TAT-CRYAB recombinant plasmid; the construction method is as follows:

[0081] S1.1. Purchase the pET28a plasmid and prepare the pET28a vector digestion system: pET28a vector, NheI, 10xFastdigestBuffer and DEPC water. The details of the pET28a vector digestion system are as follows:

[0082]

[0083] Then prepare the enzyme digestion system, bathe in a 37°C water bath for 1 hour, and then glue it back with the above steps;

[0084] S1.2, pET28a-TAT plasmid connection;

[0085] First, anneal the front and back primers of TAT, and connect after annealing. The system is: 10×T4buffer, T4 ligase, pET28a digestion product and TAT sequence annealing product.

[0086]

[0087] After the system is prepared, let it stand at room temperature for 1 hour;

[0088] S1.3, then transform the pET28a-TAT plasmid, plate, shake...

Embodiment 3

[0108] A method for applying TAT-CRYAB fusion protein in intestinal inflammation, specifically comprising the following steps:

[0109] S1. Construction of pET28a-TAT-CRYAB recombinant plasmid; the construction method is as follows:

[0110] S1.1. Purchase the pET28a plasmid and prepare the pET28a vector digestion system: pET28a vector, NheI, 10xFastdigestBuffer and DEPC water. The details of the pET28a vector digestion system are as follows:

[0111]

[0112] Then prepare the enzyme digestion system, bathe in a 37°C water bath for 1 hour, and then glue it back with the above steps;

[0113] S1.2, pET28a-TAT plasmid connection;

[0114] First, anneal the front and back primers of TAT, and connect after annealing. The system is: 10×T4buffer, T4 ligase, pET28a digestion product and TAT sequence annealing product.

[0115]

[0116] After the system is prepared, let it stand at room temperature for 1 hour;

[0117] S1.3, then transform the pET28a-TAT plasmid, plate, shake...

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Abstract

The invention discloses an application method of a TAT-CRYAB fusion protein in intestinal inflammation, and particularly relates to the field of medicines. The application method specifically comprises the following steps: S1, construction of a pET28a-TAT-CRYAB recombinant plasmid; and S2, expression and purification of the pET28a-TAT-CRYAB recombinant protein. Molecular cloning and recombinant protein technologies are mainly utilized, the pET28a-TAT-CRYAB recombinant plasmid is constructed, and the plasmid is purified to extract the recombinant protein. The application method aims to treat intestinal inflammation, body odor and treatment value in animal models are achieved, a new thought is provided for treatment of inflammatory bowel diseases, and the fusion protein can be used for treating inflammatory response induced in vitro and effectively treating intestinal inflammation of mice at present.

Description

technical field [0001] The invention relates to the technical field of medicines, and more specifically, the invention relates to an application method of a TAT-CRYAB fusion protein in intestinal inflammation. Background technique [0002] Inflammatory bowel disease (IBD) is a chronic inflammatory disease involving the digestive tract, the main manifestations are ulcerative colitis (UC) and Crohn's disease (CD). In recent years, the incidence of IBD has continued to increase, especially in developing countries. 10%-30% of UC and 70% of CD patients need surgical treatment due to drug treatment failure or complications during the course of disease development. The successful application of anti-TNF-α antibody therapy is considered a major breakthrough in the treatment of IBD, but about one-third of patients do not respond to anti-TNF-α therapy or develop drug intolerance. It is particularly urgent to study the key target genes or receptors in IBD intestinal inflammatory dise...

Claims

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Application Information

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IPC IPC(8): A61K47/64A61K38/17A61P1/00C07K19/00C12N15/62C12N15/70
CPCA61K47/645C07K14/47C12N15/70A61K38/1709A61P1/00C07K2319/10
Inventor 杜鹏许伟民
Owner 杜鹏
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